RUI: Structural and Functional Investigations of a Novel Allosteric Regulatory Mechanism in beta-Carbonic Anhydrase

RUI：β-碳酸酐酶中新型变构调节机制的结构和功能研究

• 批准号: 0741396
• 负责人: Roger Rowlett
• 金额: $ 30.15万
• 依托单位: Colgate University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0741396&HistoricalAwards=false
• 关键词: RUI; Structural; Functional; Investigations; Novel

Beta carbonic anhydrases (beta-CAs) are bicarbonate-utilizing enzymes that are essential for optimal growth in many bacteria and yeast, including some disease-causing microorganisms. Bacterial beta-CA has a unique non-catalytic bicarbonate binding site that is thought to be central to the regulation of this enzyme. This site presents a potentially attractive target without human counterpart for antimicrobial agents. The molecular basis of regulatory mechanism is not known. This research project will use methods of biophysical chemistry (including enzyme kinetics and visible spectroscopy) and structural biology (X-ray diffraction) to reveal the molecular mechanism of this regulatory site. This knowledge could be exploited in the future to design molecules to specifically interfere with the function of this enzyme for anti-microbial purposes, or to gain insight into the metabolic role of the enzyme in bacteria and other microorganisms in which it is present. There are several broader impacts of the research project, one of which is the involvement of undergraduate students in research projects that provide training in advanced, current praxis in protein chemistry and structural biology. A high percentage of these students are expected to enter into advanced degree programs in the physical and life sciences, and ultimately the domestic scientific work force. In addition, a post-doctoral fellow with an interest in entering the undergraduate education community will be trained in biophysical chemistry and structural biology. Written laboratory guides appropriate for involving undergraduates or other novice investigators in modern praxis in protein engineering and X-ray crystallography will be updated and freely disseminated to the scientific community. Finally, the funding of critical research equipment replacement and upgrades will significantly enhance the biophysical chemistry research infrastructure at Colgate University.

β-碳酸酐酶（β-CA）是一种利用碳酸氢盐的酶，对于许多细菌和酵母（包括一些致病微生物）的最佳生长至关重要。细菌β-CA具有独特的非催化碳酸氢盐结合位点，该位点被认为是该酶调节的中心。 该位点提供了一个潜在的有吸引力的靶点，而没有抗微生物剂的人类对应物。调控机制的分子基础尚不清楚。本研究将利用生物物理化学（包括酶动力学和可见光谱）和结构生物学（X射线衍射）的方法来揭示该调控位点的分子机制。这一知识可以在未来被利用来设计分子，以特异性地干扰这种酶的功能，用于抗微生物目的，或者深入了解这种酶在细菌和其他微生物中的代谢作用。该研究项目有几个更广泛的影响，其中之一是本科生参与研究项目，提供蛋白质化学和结构生物学先进的，当前的实践培训。预计这些学生中有很大一部分将进入物理和生命科学的高级学位课程，并最终成为国内科学工作者。此外，一名有兴趣进入本科教育界的博士后研究员将接受生物物理化学和结构生物学方面的培训。将更新适合于本科生或其他新手研究人员参与蛋白质工程和X射线晶体学现代实践的书面实验室指南，并免费分发给科学界。最后，关键研究设备更换和升级的资金将大大加强高露洁大学的生物物理化学研究基础设施。